环介导等温扩增核酸技术及其应用

Loop-mediated Isothermal Amplification Method for Detection of Nucleic Acids and its Application

摘要:

环介导等温扩增（loop-mediated isothermal amplification，简称 LAMP）是利用4个特殊设计的引物和具有链置换活性的DNA聚合酶，在恒温条件下特异、高效、快速地扩增DNA的新技术。该技术在1h内能扩增出10⁹靶序列拷贝，扩增产物是一系列反向重复的靶序列构成的茎环结构和多环花椰菜样结构的DNA片段的混合物，电泳后在凝胶上显现出由不同大小的区带组成的阶梯式图谱。LAMP技术以其特异性强、灵敏度高、快速、准确和操作简便等优点在核酸的科学研究、疾病的诊断和转基因食品检测等领域得到了日益广泛的应用。

Abstract:

A novel nucleic acid amplification method, termed loop-mediated isothermal amplification (LAMP), which amplifies DNA with high specificity, efficiency, and rapidity under isothermal conditions, may be a valuable tool for the rapid detection of infectious diseases. This method employs a DNA polymerase that have activity of strand displacement DNA synthesis and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA. LAMP can amplify a few copies of DNA to 10⁹ in less than an hour. The final products are stem-loop DNA with several inverted repeats of the target and cauliflower-like structures with multiple loops. A positive reaction would be shown as a ladder-like pattern in a gel electrophoresis analysis. Because of the advantage, the LAMP method will be widely applied to research of nucleic acid, clinical diagnosis of infectious diseases and detection of genetically modified organisms etc.